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DNA Isolation Lab

DNA Isolation Lab. Jim Evans MD, Ph.D ASTAR Judicial Conference Chapel Hill, NC 16 March 2011. Understanding Genetics. Heredity. by Thomas Hardy. 1840-1928. The years-heired feature that can In curve and voice and eye Despise the human span Of durance - that is I;

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DNA Isolation Lab

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  1. DNA Isolation Lab Jim Evans MD, Ph.D ASTAR Judicial Conference Chapel Hill, NC 16 March 2011

  2. Understanding Genetics Heredity by Thomas Hardy 1840-1928 The years-heired feature that can In curve and voice and eye Despise the human span Of durance - that is I; The eternal thing in man, That heeds no call to die. I Am the family face; Flesh perishes, I live on, Projecting trait and trace Through time to times anon, And leaping from place to place Over oblivion

  3. Understanding Genetics Through the Application of Science • Gregor Mendel • Rediscovery of Mendel’s Laws • Formulation of the chromosomal basis of inheritance 1864-1933 1848-1935 1822-1884 1890-1967

  4. The Material Gene • Friedrich Miescher 1869 • Fredrick Griffith 1928 • Oswald Avery et al. 1943 • Watson, Crick, Franklin 1953 • The Human Genome Project 2000

  5. IMPRESS YOUR FRIENDS! IT’S FUN! Isolate Your Own DNA IT’S 100% SAFE! IT’S EASY!

  6. Swish the Gatorade • Each participant will get a small cup with 2ml of Gatorade. • To get buccal cells (cells which line the inside of the cheek) • Swish the Gatorade in your mouth for 1 minute while scraping the inside of your cheeks with your teeth • Do not gargle, swallow or spit on the instructor

  7. Spit and Pour • Spit the Gatorade back into the cup • Pour the Gatorade from the cup into the blue-topped tube

  8. Add “Lysing” solution • Add lysing solution to the blue top tube and invert several times to mix well • Lysing solution breaks open the cell membrane and liberates the cell content, including the DNA • The solution will be viscous and trap air bubbles

  9. Add Alcohol • Add a pipetteful of Alcohol and wait a couple of minutes. You should see a white stringy clump arising from the water-alcohol interface • That is your DNA! • DNA is not soluble in alcohol so it precipitates as a white clump

  10. Retrieve your DNA • Pick the DNA clump up with a pipette and transfer to a small tube to use as a necklace if you desire • If you did not get anything: • You did not swish long enough • You can try again and repeat all the steps • You are a non-DNA based alien organism. We will be sending the sample to the NSA for further analysis and you may be visited soon by Federal Agents

  11. Why is This Science? • Reproducible • Amenable to prediction and verification • Amenable to experiment • Independent of ideology • Amenable to revision Does it consistently work and can it possibly be improved upon?

  12. Is This Science? MARCH 04, 2011 Anthony Atala printed a functioning kidney on stage at the TED conference Anthony Atala, printed a biocompatible model of a human kidney on stage at the 2011 TED conference Thursday, in a technique that could someday be used to create new organs from a patient’s own tissue rather than relying on donated organs.Atala explained the process whereby scanners build a 3-D image of the kidney, then a tissue sample smaller than a postage stamp was used to seed the process. The organ printer then built the kidney layer-by-layer, creating an almost perfect replica of the patient’s tissue. http://nextbigfuture.com/2011/03/anthony-atala-printed-functioning.html

  13. What is DNA and Why Does it Matter? Molecular Biology in 10 Slides

  14. All living things are composed of cells: Skin Brain Liver Blood etc……….

  15. antibodies channels nucleus cytoplasm Structural components Tiny machines made of protein carry out the functions of life…

  16. At the Center of Each Cell Lies the Nucleus, containing The DNA Double Helix Sugar phosphate backbone Base pair Bases Adenine (A) Thymine (T) Cytosine (C) Guanine (G) ASCO

  17. DNA DNA has two jobs in the living organism… It directs the synthesis of Proteins It serves as a store of information which can be transmitted to future generations

  18. DNA’s First Job: Directing Protein Synthesis DNA is every cell’s “instruction book” DNA directs the synthesis of proteins The order of nucleotides (DNA’s basic building blocks) dictates the order of amino acids in protein chains The order of amino acids in a protein dictates how that protein chain will fold and therefore what its structure and function will be (i.e. the shape of the protein “machine”)

  19. Gene #1 Gene #2 Gene #3 A Gene is simply a segment of DNA that directs the synthesis of one particular protein...

  20. DNA’s Second Job • DNA serves as a store of information • Passing on our genetic information from one cell to its daughter cells or from one generation to another How does DNA manage to pass its information on to the next generation?

  21. The DNA Double Helix Sugar phosphate backbone Base pair Bases Adenine (A) Thymine (T) Cytosine (C) Guanine (G) ASCO

  22. Passing Along the Information By virtue of being a double helix, each DNA molecule contains a copy of itself To make a new cell (or a sperm or egg and thus the next generation), the cell separates the two strands of DNA and copies each, forming a new identical double helix One copy is sent to each “daughter” cell Thus, each new cell (or new generation) receives the exact same information necessary for propagation of life

  23. T T T T T T C C C C C C A A A A A A T T T T T T G G G G G G A A A A A A C C C C C C G G G G G G A A G G T T A A C C T T G G A A A A G G G G T T T T A A A A C C C C T T T T G G G G C C C C C C The cell divides and sends one copy of each double helix to each new cell Copies of each strand are made The double helix is unzipped

  24. Molecular Biology in 1 Slide Gene Nucleus Cell Chromosomes Protein Adapted from Understanding Gene Testing, NIH, 1995 ASCO

  25. Human Genome “There’s a gene in here that explains why you’re such an idiot”

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